Rapid Detection of β-Lactamase-Hydrolyzing Extended-Spectrum Cephalosporins in Enterobacteriaceae by Use of the New Chromogenic βLacta Test

Morosini, María Isabel; García-Castillo, María; Tato, Marta; Gijón, Desirée; Valverde, Aránzazu; Ruíz-Garbajosa, Patricia

doi:10.1128/jcm.03614-13

Cited by 25 publications

(17 citation statements)

References 19 publications

(24 reference statements)

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“…The ␤-Lacta test, which is designed to detect those AmpC overproducers, failed to detect those strains in 10/14 of the cases. A similar failure of detection was reported by Morosini et al (13). The overall specificity of the ␤-Lacta test for detecting ESBL producers was much lower than that of the Rapid ESBL NDP test, since it also partially detects AmpC producers and several types of carbapenemase producers.…”

Section: Discussionmentioning

confidence: 70%

Comparison of Three Biochemical Tests for Rapid Detection of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae

2016

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dEnterobacterial isolates producing clavulanic-inhibited extended-spectrum ␤-lactamases (ESBLs) are increasingly spreading in the community and are often responsible for nosocomial infections. Rapid biochemical tests have been developed recently for their detection. Three tests, namely, the Rapid ESBL NDP test, the ␤-Lacta test, and the Rapid ESBL Screen, have been evaluated with a collection of 108 well-characterized strains, including wild-type strains, strains producing ESBLs, overexpressed cephalosporinases, and carbapenemases. The ESBL NDP test and the Rapid ESBL Screen (a copy of the ESBL NDP test) are aimed at detecting ESBL producers, while the ␤-Lacta test is aimed at detecting not only ESBL producers but also cephalosporinase and carbapenemase producers. The sensitivity and specificity for detecting ESBL producers (n ‫؍‬ 60) were 95% and 100% for the Rapid ESBL NDP test, 80% and 87% (after 30 min) and 92% and 83% (after 2 h) for the Rapid ESBL Screen, and 88% and 71% for the ␤-Lacta test, respectively. Varied and time-consuming detection (up to 2 h) of ESBLs by the Rapid ESBL Screen and concomitant and varied detection of producers of AmpC and several types of carbapenemases correspond to significant shortcomings of using the Rapid Screen ESBL and ␤-Lacta tests, respectively.A cquired resistance to broad-spectrum cephalosporins in Enterobacteriaceae is mainly due to the production of clavulanic acid-inhibited extended-spectrum ␤-lactamases (ESBLs), which have extensively disseminated worldwide (1). Along with carbapenemase producers, ESBL producers represent the most important resistance trait in Enterobacteriaceae in 2015. The European Antimicrobial Resistance Surveillance System Network (EARSNet), which includes 30 European countries, reported in 2013 the prevalence rates of nonsusceptibility to broad-spectrum cephalosporins among invasive enterobacterial isolates (2). The proportion of Escherichia coli strains resistant to broad-spectrum cephalosporins ranged from 5% to 39.6%, depending on the country. In Klebsiella pneumoniae, the percentage of resistance to broad-spectrum cephalosporins showed a significant increase from 22.8% in 2012 to 30% in 2013, encompassing 85 to 100% of the ESBL producers. In the United States, the percentages of health care-associated infections caused by broad-spectrum cephalosporin-resistant Enterobacteriaceae have been estimated to be 14% and 23% for E. coli and Klebsiella spp., respectively (http://www.cdc.gov /drugresistance/threat-report-2013). Those ESBL-producing Enterobacteriaceae are identified either as a source of hospital-or community-acquired infections (3).The rapid detection of ESBL producers is therefore crucial in order to prevent their dissemination and to guide the treatments of infected patients. Several phenotypic techniques are based on the inhibition of ESBL activity by clavulanic acid or tazobactam. Those techniques require a preliminary growth step of 24 to 48 h (4). Molecular detection of ESBL-coding genes is interesting but remains costly, requi...

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Section: Discussionmentioning

confidence: 70%

Comparison of Three Biochemical Tests for Rapid Detection of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae

2016

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“…The protocol of the ESBL NDP test first devised with cultured bacteria has been modified here and led to a shorter period of detection, which was reduced from 60 min (5) to 15 min. Another chromogenic test called the Cica-␤-test or ␤Lacta test (Bio-Rad) has been recently developed for the rapid (15-min) detection of Enterobacteriaceae resistant to expanded-spectrum cephalosporins from cultures grown on agar media (12)(13)(14). A prospective evaluation performed with the ␤Lacta test on enterobacterial isolates found sensitivity, specificity, PPV, and NPV values of 87.7%, 99.6%, 97.1%, and 98.2%, respectively (13).…”

Section: Discussionmentioning

confidence: 99%

“…The most recent study published using the ␤Lacta test reported on false-negative results obtained with AmpC overproducers, producers of ESBL at low levels, and producers of VIMtype carbapenemase (14). In addition, the ␤Lacta test has not been evaluated for detection of ESBL-E directly from clinical samples.…”

Section: Rapid Detection Of Esbl Producers From Urinementioning

confidence: 99%

Rapid Detection of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae from Urine Samples by Use of the ESBL NDP Test

2014

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From June to September 2012, 500 urine samples were recovered from patients with urinary tract infections (UTI) due to Gramnegative bacilli (>10 4 leukocytes/ml and >10 5 Gram-negative isolates/ml) who visited the University hospital Bicêtre (France). They were challenged with extended-spectrum-␤-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-E) using the rapid diagnostic ESBL NDP test. Results of the ESBL NDP test were compared to the results of the double-disc susceptibility test (DDST) performed on solid-agar plates and molecular identification of the ␤-lactamase genes. Among the 450 nonduplicate urine samples, 11.3% were positive for ESBL-E by using the DDST, the ESBL determinants being mostly of the CTX-M type (CTX-M-15) according to molecular testing. Results of the ESBL NDP test were obtained within 15 min. The sensitivity and specificity of the ESBL NDP test were 98% and 99.8%, respectively, whereas the positive and negative predictive values of this test were 98% and 99.8%, respectively. A perfect correlation between cefotaxime resistance and positivity of the ESBL NDP test was observed. Therefore, the ESBL NDP test offers a powerful tool for a rapid identification of ESBL-E and associated resistance to expandedspectrum cephalosporins. It may be useful in particular for guiding first-line antibiotic therapy.

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“…However, the fact that hydrolysis of cefotaxime is not complete can explain why OXA-48-producing enterobacterial isolates remain phenotypically susceptible to cefotaxime in vitro. Activity on cefotaxime can also explain the eventual positive results obtained by colorimetric assays based on pH reduction, aimed at ESBL detection [15,16], which could be due to carboxyl-acid formation after hydrolysis of the beta-lactam ring. From a clinical point of view, the efficacy of cefotaxime against OXA-48-producing Enterobacteriaceae could be impaired by the aforementioned hydrolysis and this is consistent with the previously reported lower efficacy of this drug in vivo with respect to ceftazidime or cefepime [8,9].…”

Section: Resultsmentioning

confidence: 99%

Analysis of the Degradation of Broad-Spectrum Cephalosporins by OXA-48-Producing Enterobacteriaceae Using MALDI-TOF MS

et al. 2019

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The objective of the study was to evaluate the activity of OXA-48 against different broad-spectrum cephalosporins and to identify the reaction products by MALDI-TOF MS. The action of OXA-48 on cefotaxime, ceftazidime, and ceftriaxone was assessed by this method, using an Escherichia coli J53 transconjugant carrying only the ~62 Kb IncL plasmid containing the blaOXA-48 gene, and the same strain without any plasmid was included as a negative control. In addition, a collection of 17 clinical OXA-48-producing Enterobacteriaceae, which were susceptible to broad-spectrum cephalosporins, was evaluated. MALDI-TOF MS-based analysis of the E. coli transconjugant carrying the blaOXA-48-harboring plasmid, and also the clinical isolates, showed degradation of cefotaxime into two inactive compounds—decarboxylated and deacetylated cefotaxime (~370 Da) and deacetyl cefotaxime (~414 Da), both with the hydrolyzed beta-lactam ring. Reaction products were not obtained when the experiment was performed with ceftriaxone or ceftazidime. From a clinical point of view, our study supports the idea that the efficacy of cefotaxime against OXA-48-producing Enterobacteriaceae is doubtful, in contrast to ceftazidime and ceftriaxone which could be valid choices for treating infections caused by these bacteria. However, further clinical studies confirming this hypothesis are required.

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Rapid Detection of β-Lactamase-Hydrolyzing Extended-Spectrum Cephalosporins in Enterobacteriaceae by Use of the New Chromogenic βLacta Test

Cited by 25 publications

References 19 publications

Comparison of Three Biochemical Tests for Rapid Detection of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae

Comparison of Three Biochemical Tests for Rapid Detection of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae

Rapid Detection of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae from Urine Samples by Use of the ESBL NDP Test

Analysis of the Degradation of Broad-Spectrum Cephalosporins by OXA-48-Producing Enterobacteriaceae Using MALDI-TOF MS

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